EP1368364B1 - C2, 8-disubstituted adenosine derivatives and their different uses - Google Patents

C2, 8-disubstituted adenosine derivatives and their different uses Download PDF

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EP1368364B1
EP1368364B1 EP02700559A EP02700559A EP1368364B1 EP 1368364 B1 EP1368364 B1 EP 1368364B1 EP 02700559 A EP02700559 A EP 02700559A EP 02700559 A EP02700559 A EP 02700559A EP 1368364 B1 EP1368364 B1 EP 1368364B1
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compound
alkyl
group
formula
alkylamine
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French (fr)
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EP1368364A1 (en
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Erica Van Tilburg
Ad Ijzerman
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Can Fite Biopharma Ltd
Universiteit Leiden
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Can Fite Biopharma Ltd
Universiteit Leiden
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives

Definitions

  • the present invention relates to novel C2,8-disubstituted adenosine derivatives, their process of preparation and different uses.
  • Adenosine mediates a wide variety of effects as a result of its activation of specific membrane-bound receptors called P 1 -purinoceptors.
  • the three subclasses of P 1 -purinoceptors are A 1 , A 2 and A 3 , with A 2 further subdivided into A 2A and A 2B .
  • All adenosine receptors are coupled to the enzyme adenylate cyclase; activation of the A 1 and A 3 adenosine receptors inhibit the adenylate cyclase, whereas activated A 2A and A 2B receptors stimulate it.
  • the adenosine A 2A receptor can be found throughout the whole body, and A 2A receptor agonists might be used to inhibit platelet aggregation in thrombosis, in the diagnosis of diseases in coronary arteries, in ischemia and reperfusion. 1 Furthermore, activation of adenosine A 2A receptors has been shown to alter the binding characteristics of other receptors. Stimulation of adenosine A 2A receptors in rat striatal membranes reduces the affinity of agonist binding to dopamine D 2 receptors. 2,3 This raises the possibility of using adenosine A 2A receptor agonists as a novel therapeutic approach in the treatment of psychosis.
  • partial agonists for the adenosine A 2A receptor may be accompanied with serious side effects such as cardiovascular actions, since the receptor is ubiquitously distributed.
  • the design of partial agonists for the adenosine receptors has already been shown to be a useful tool to achieve selectivity of action in vivo by exploitation of the differences in receptor-effector coupling in various tissues. 4-7
  • partial agonists for the adenosine A 2A receptor might then be developed as antipsychotic agents also devoid of undesired cardiovascular actions.
  • Selectivity for the adenosine A 2A receptor can be obtained by the introduction of a C2-substituent, such as a 1-hexynyl or a 1-hexenyl group that have been shown to induce high affinity for the adenosine A 2A receptor compared to A 1 .
  • a C2-substituent such as a 1-hexynyl or a 1-hexenyl group that have been shown to induce high affinity for the adenosine A 2A receptor compared to A 1 .
  • Partial agonism for adenosine receptors in general has been achieved by the introduction of alkylthio-substituents at the 5'-position. 5,11 However, partial agonism for the adenosine A 1 receptor has also successfully been accomplished by introducing alkylamino substituents at the C8-position of adenosine. 12 8-Alkylamino substituted CPA derivatives have proven to be adenosine
  • US 5,877,180 discloses agonists of A 2A receptors for the treatment of inflammatory diseases.
  • WO 00/78777 discloses propargyl phenyl ether A 2A receptor agonists.
  • the present invention provides C2,8-disubstituted adenosine derivatives having the following general formula (I) (hereinafter referred to at times as the "the compound of the present invention ”): wherein
  • lower alkyl refers to any saturated hydrocarbon, either linear or branched chain comprising from one to ten carbon atoms in the backbone of the hydrocarbon.
  • lower alkenyl and lower alkynyl refer to a linear or branched hydrocarbon comprising from two to ten carbon atoms in the backbone, wherein at least two of the carbon atoms are connected via a double or triple bond, respectively.
  • the compound of the invention also includes salts of the compound of formula (I) and in particular, physiologically acceptable salts.
  • physiologically acceptable salt refers to any non-toxic alkali metal, alkaline earth metal, and ammonium salts commonly used in the pharmaceutical industry, including the sodium, potassium, lithium, calcium, magnesium, barium ammonium and protamine zinc salts, which are prepared by methods known in the art.
  • non-toxic acid addition salts which are generally prepared by reacting the compounds of this invention with a suitable organic or inorganic acid.
  • the acid addition salts are those which retain the biological effectiveness and properties of the free bases and which are not biologically or otherwise undesirable.
  • acids are those derived from mineral acids, and include, inter aila, hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, metaphosphoric and the like.
  • Organic acids include, inter alia, tartaric, acetic, propionic, citric, malic, malonic, lactic, fumaric, benzoic, cinnamic, mandelic, glycolic, gluconic, pyruvic, succinic salicylic and arylsulphonic, e.g. p-toluenesulphonic, acids.
  • the present invention also provides a process for the preparation of the compound of the present invention (i.e. the compound of formula (I) as defined above) or a salt thereof, the process comprising in general treating a compound of the formula (II): wherein
  • the process of the present invention may provide a compound wherein R 2 and are R 3 are the same or different, depending on the amount and number of reagents employed.
  • the process is preferably carried out in two steps, each step comprising treatment with 1 equivalent of one of said reagents carrying different substituents.
  • the invention also provides pharmaceutical compositions comprising as active ingredient an effective amount of one or more of the compounds of the invention as defined hereinbefore.
  • the present invention is based on the surprising finding that C8-substituted and C2,8-disubstituted adenosine derivatives display higher adenosine A 2A receptor affinity and/or A 2A selectivity compared to the A 1 and A 3 receptors.
  • the present invention provides C2,8-disubstituted adenosine derivatives having the following general formula (I): wherein
  • the substituent R 2 represents a halogen atom, a C 2 -C 10 alkenyl or C 2 -C 10 alkynyl or (ar)alkylamine. More preferably, the alkenyl or alkynyl groups are, respectively C 6 -alkenyl or C 6 -alkynyl, the halogen atom is iodine and the (ar)alkylamine group is benzylamine.
  • R 2 is a halogen atom, preferably an iodine atom.
  • R 2 is 1-hexenyl or 1-hexynyl, the former preferably representing the ( E ) 1-hexenyl isomer.
  • the R 3 substituent preferably represents an alkylamine, aralkylamine, alkynyl or alkynyl group.
  • the alkylamine is preferably selected from methylamine, ethylamine, propylamine, butylamine, the alkynyl is prefereably 1-hexynyl, and the (ar)alkylamine is preferebly benzylamine, however the latter may also include anilines and substituted arylamines.
  • biologically active indicates that the compound of the present invention has some sort of a biological activity, for example, a measurable effect on a target receptor.
  • the compound of the present invention may induce adenosine receptor activity, preferably acting as adenosine receptor agonists and more preferably as adenosine A 2A receptor agonists.
  • agonist refers to a biologically active ligand, which binds to its complementary biologically (active) receptor and activates the latter either to cause a biological response in the receptor or to enhance preexisting biological activity of the receptor.
  • the agonist in fact mimics the effect of the natural ligand, in the present case, adenosine, or at times, even increases or prolongs the duration of the biological effect obtained as compared to the effect induced by the natural ligand.
  • the compounds of the present invention were specifically found to activate adenosine receptors, with a higher selectivity and affinity for the adenosine A 2A receptor.
  • the compounds of the present invention may be recognized as adenosine A 2A receptor agonists. More preferably and as will be shown in the following Specific Examples, the compounds of the present invention are partial agonists of the adenosine A 2A receptor. receptor if it produces (or induces (e.g. when increased in concentration) the maximal possible response achievable by activation of this receptor.
  • a compound according to the invention is considered a " partial agonist” if it is unable to produce maximal activation of the receptor to which it binds no matter how high is its concentration.
  • Preferred compounds according to the present invention include:
  • the present invention provides a process for the preparation of a compound of the general formula (I): in which
  • the process of the invention may be carried out in a single step reaction, according to winch two or more equivalents of the reagent selected are applied on the starting compound, or in two steps.
  • the resulting compound may contain the same or different substituents in R 2 and R 3 depending on the amount of reagent employed.
  • a compound of the following general formula (III) is obtained in which only C8 is substituted, while when using two or more equivalents of the reagent a compound in which C2 and C8 are substituted with the same group is obtained.
  • the starting compound of formula (II) employed by the process of the present invention may be obtained, for example, by reacting with bromine, in the presence of a base, a compound of the following formula (V): in which R 1 are the same or different and represent a hydrogen atom or a methylcarbonyl group.
  • a compound of the following formula (V): in which R 1 are the same or different and represent a hydrogen atom or a methylcarbonyl group in which R 1 are the same or different and represent a hydrogen atom or a methylcarbonyl group.
  • the compounds obtained by the process of the invention are preferably those in which R 2 represents a halogen atom, a lower alkenyl or lower alkynyl and R 3 is an alkylamine, (ar)alkyl amine or alkynyl.
  • R 2 is iodine or a C 6 -alkenyl or C 6 -alkynyl group, the latter being respectively, in preference, 1-hexenyl or 1-hexynyl, the 1-hexenyl group being preferably the ( E ) 1-hexenyl isomer.
  • R 3 is an alkylamine group, it is preferably a lower alkylamine selected from the group consisting of methylamine, ethylamine, propylamine, butylamine, while in case R 3 is an (ar)alkylamine it is in preference benzylamine.
  • R 2 is 1-hexynyl. Yet, according to another embodiment, R 2 is ( E ) 1-hexenyl. In both embodiments R 3 is preferably selected from methylamine, ethylamine, propylamine and butylamine.
  • the compound 2-(1-Hehynyl)adenosine ( 3 ) was prepared (in good yield (85%)) by reacting 2-iodoadenosine ( 1 ) with 1-hexyn. 8,15
  • Compound 4 was prepared by reacting 1 with ( E )-1-(borocatechol)-1-hexene. 10
  • unprotected 2-iodoadenosine ( 1 ) or 2',3',5'-tri- O -acetyl-2-iodoadenosine ( 2 ) was brominated at the C8-position.
  • the present invention provides pharmaceutical compositions comprising as active ingredient a therapeutically effective amount, including a prophylactically effective amount, of one or more of the compounds of formula (I), or a salt of said compound, and a pharmaceutically acceptable additive.
  • a therapeutically effective amount including a prophylactically effective amount, of one or more of the compounds of formula (I), or a salt of said compound, and a pharmaceutically acceptable additive.
  • Preferred compounds employed in the compositions of the present invention are as defined above.
  • an effective amount for the purposes described herein is that determined by such considerations as are known to those versed in the art.
  • the amount must be sufficient to achieve a desired therapeutic effect, e.g. to treat a disease or a disorder.
  • the effective amount is typically determined in appropriately designed clinical trials (e.g. dose range studies) and the person versed in the art will know how to properly conduct such trials in order to determine the effective amount.
  • an effective amount depends on a variety of factors including the affinity of the ligand to the receptor, its distribution profile within the body, a variety of pharmacological parameters such as half life in the body, on undesired side effects, if any, on factors such as age and gender, etc.
  • compositions of the invention are preferably for the treatment of diseases associated with the function of adenosine A 2A receptor.
  • treatment refers to the administering of a therapeutic amount of th e compound or composition of the present invention which is effective to ameliorate undesired symptoms associated with a disease, to prevent the manifestation of such symptoms before they occur, to slow down the progression of a disease, to slow down the deterioration of symptoms, to slow down the irreversible damage caused by the chronic stage of a disease, to lessen the severity or cure a disease, to improve survival rate or more rapid recovery, to prevent the disease from occurring, or a combination of two or more of the above.
  • Treatment according to the invention preferably includes activation by one or more of the compounds of the present invention of an adenosine receptor (or elevation of an already active receptor) and preferably of the adenosine A 2A receptor.
  • additives used herein refers to any substance combined with said compound and include, without being limited thereto, diluents, excipients, carriers, solid or liquid fillers or encapsulating materials which are typically added to formulations to give them a form or consistency when it is given in a specific form, e.g. in pill form, as a simple syrup, aromatic powder, and other various elixirs.
  • the additives may also be substances for providing the formulation with stability, sterility and isotonicity (e.g. antimicrobial preservatives, antioxidants, chelating agents and buffers), for preventing the action of microorganisms (e.g. antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid and the like) or for providing the formulation with an edible flavor etc.
  • the additives are inert, non-toxic materials, which do not react with the active ingredient of the invention.
  • the additives may be designed to enhance the binding of the active agent to its receptor.
  • the term additive may also include adjuvants, being substances affecting the action of the active ingredient in a predictable way.
  • the additives can be any of those conventionally used and are limited only by chemico-physical considerations, such as solubility and lack of reactivity with the compound of the invention, and by the route of administration.
  • the active agent of the invention is preferably to be administered orally to the patient.
  • Conventional methods such as administering the compound/s in tablets, suspensions, solutions, emulsions, capsules, powders, syrups and the like are usable.
  • composition of the invention may contain additives for facilitating oral delivery of the compound/s of the invention.
  • Formulations suitable for oral administration can consist of (a) liquid solutions, such as an effective amount of the compound dissolved in diluents, such as water, saline, or orange juice; (b) capsules, sachets, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient, as solids or granules; (c) powders; (d) suspensions in an appropriate liquid; and (e) suitable emulsions.
  • Liquid formulations may include diluents, such as water and alcohols, for example, ethanol benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent, or emulsifying agent
  • diluents such as water and alcohols, for example, ethanol benzyl alcohol
  • polyethylene alcohols either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent, or emulsifying agent
  • Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and corn starch.
  • Tablet forms can include one or more of lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodiumk talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible carriers.
  • Lozenge forms can comprise the active agent in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like.
  • an inert base such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like.
  • Such additives are known in the art.
  • the compound/s may to be administered to the patient parenterally.
  • the composition will generally be formulated in a unit dosage injectable form (solution, suspension, emulsion).
  • Pharmaceutical formulation suitable for injection may include sterile aqueous solutions or dispersions and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • the carrier can be a solvent or dispersing medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, lipid polyethylene glycol and the like), suitable mixtures thereof and vegetable oils such as cottonseed oil, sesame oil, olive oil, soybean oil, corn oil, sunflower oil, or peanut oil; a fatty acid esters such as ethyl oleate and isopropyl myristate and variety of other solvent systems as known per se.
  • the carrier may be chosen based on the physical and chemical properties of the active agent.
  • the active ingredient has a poor water solubility, and an oily carrier is therefore used proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.
  • Suitable soaps for use in parenteral formulations include fatty alkali metal, ammonium, and triethanolamine salts
  • suitable detergents include (a) cationic detergents such as, for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides, (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxy- ethylenepolypropylene copolymers, (d) amphoteric detergents such as, for example, alkyl- ⁇ -aminopriopionates, and 2-alkyl imidazoline quaternary ammonium salts,
  • compositions may contain one or more nonionic surfactants having a hydrophile-lipophile balance (HLB) of from about 12 to about 17.
  • Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol.
  • composition of the present invention may include one or more of the compounds of the present invention and may be comprise other biologically active substances, to provide a combined therapeutic effect.
  • the compounds of the present invention were found to be capable of binding and activating or inducing the activity of adenosine receptors, particularly, of the adenosine A 2A receptor.
  • the composition of the present invention is preferably for the treatment of a disease or a disorder, which require for their treatment activation or induction of the activity of the adenosine A 2A receptor present on a target cell.
  • the compound or composition of the present invention may be useful as therapeutic or prophylactic agents for circulatory diseases, such as hypertension and ischemic heart or brain disease.
  • the compounds or compositions of the present invention may be useful as neuroleptic agents, e.g. for the treatment of psychosis, or for wound healing.
  • compositions of the present invention as set forth hereinabove and below are administered and dosed in accordance with good medical practice, taking into account the clinical conditions of the individual patient, the site and method of administration, scheduling of administration, individual's age, sex body weight and other factors known to medical practitioners.
  • the dose may be single daily dose or multiple doses over period of several days.
  • the treatment generally has a length proportional to the length of the disease process and drug effectiveness and the individual species being treated Suitable doses and dosage regimens can be determined by conventional range-finding techniques known to those of ordinary skill in the art. Generally, treatment is initiated with smaller dosages, which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments, until the optimum effect under the is reached. Exemplary dosages range from about 0.001 mg/kg body weight to about 10 mg/kg body weight of the subject being treated/day.
  • 2-Iodoadenosine (comparative compound 1) was prepared according to literature. 17 Yeld 80%; mp 185-187 ⁇ C; Rf 0.21 (10% MeOH in CH 2 Cl 2 ).
  • 2-iodo-8-bromoadenosine (Comparative compound 5).
  • 2-Iodoadenosine (1, 2.93 g, 7.45 mmol) was dissolved in NaOAc buffer (1.0 M, pH 4, 50 ml) at 50 °C. The solution was cooled to room temperature and bromine (0.46 mL, 8.94 mmol) was added. After stirring overnight at room temperature, adding NaHSO 3 destroyed the excess of bromine and the pH of the solution was adjusted to 7 with NaOH solution (5 M). The reaction mixture was kept at 4 °C for 5hr and the precipitate was filtered. The white solid was washed with water and dried.
  • Amination of compound 5 or 6 to obtain the 8-alkylamino-2-iodoadenosines 7-10 was generally as follows: to 8-bromo-2-iodoadenosine (5) or 2',3',5'-tri- O -acetyl-2-iodo-bromoadenosine ( 6 ) (0.17 mmol) the appropriate alkylamine was added (excess) and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated in vacuo and the product was crystallised from water.
  • 2-iodo-8-ethylaminoadenosine (compound 8).
  • the reaction was performed with 2-iodo-8-bromoadenosine ( 5 , 90 mg, 0.19 mmol) and ethylamine (2.5 ml, 70% w/v in water).
  • 2-Iodo-8-propylaminoadenosine (compound 9): The reaction was performed with 2-iodo-8-bromoadenosine ( 5 , 90 mg, 0.19 mmol) and propylamine (2.5 ml, 70% w/v in water).
  • 2-Iodo-8-butylaminoadenosine (compound 10): The reaction was performed with 2-iodo-8-bromoadenosine ( 5 , 1.15 g, 2.44 mmol) and n- butylamine (25 ml). Some drops of water were added to dissolve everything.
  • 2-iodo-8-benzylaminoadenosine (compound 11): 8-Bromo-2-iodoadenosine ( 5 , 1.28 g, 2.14 mmol) was dissolved in benzylamine (21.4 mmol, 2.34 ml) and some drops of water were added to dissolve everything. The mixture was stirred overnight at 60 °C and concentrated in vacuo. The white solid was stirred in CH 2 Cl 2 , filtered and dried.
  • assays were performed in 50/10/1 buffer (50 mM Tris/10mM MgCl 2 /1mM ethylenediaminetetra-acetic acid (EDTA) and 0.01% 3-([3-cholamidopropyl]-dimethylammonio)-1-propanesulfonate (CHAPS)) in glass tubes and contained 50 ⁇ l of a HEK 293 cell membrane suspension (10-30 ⁇ g), 25 ⁇ L [ 125 I]AB MECA (final concentration 0.15 nM), and 25 ⁇ l of ligand.
  • 50/10/1 buffer 50 mM Tris/10mM MgCl 2 /1mM ethylenediaminetetra-acetic acid (EDTA) and 0.01% 3-([3-cholamidopropyl]-dimethylammonio)-1-propanesulfonate (CHAPS)
  • Incubations were carried out for 1 hr at 37°C and were terminated by rapid filtration over Whatman GF/B filters, using a Brandell cell harvester (Brandell, Gaithersburg, MD). Tubes were washed three times with 3 ml of buffer. Radioactivity was determined in a Beckman 5500B ⁇ -counter. Nonspecific binding was determined in the presence of 10 -5 M R-PIA.
  • CHO cells expressing the human adenosine A 2A receptors were grown overnight as a monolayer in 24 wells tissue culture plates (400 ⁇ l/well; 2x10 5 cells/well).
  • cAMP generation was performed in Dulbecco's Modified Eagles Medium (DMEM)/N-2-hydroxyethylpiperazin-N'-2-ethansulfonic acid (HEPES) buffer (0.60 g HEPES/50 ml DMEM pH 7.4).
  • DMEM Dulbecco's Modified Eagles Medium
  • HEPES N-2-hydroxyethylpiperazin-N'-2-ethansulfonic acid
  • DMEM/HEPES buffer 250 ⁇ l
  • 100 ⁇ l DMEM/HEPES buffer 100 ⁇ l adenosine deaminase (final concentration 5 IU/ml) and 100 ⁇ l of a mixture of rolipram and cilostamide (final concentration 50 ⁇ M each) were added.
  • 100 ⁇ L agonist was added.
  • the reaction was terminated by removing the medium and adding 200 ⁇ l 0.1 M HCl.
  • Wells were stored at -20°C until assay.
  • cAMP cAMP binding protein 12
  • a buffer 150 mM K 2 HPO 4 / 10 mM EDTA / 0.2% Bovine Serum Albumine (BSA) at pH 7.5.
  • Samples (20 ⁇ l + 30 ⁇ l 0.1 M HCl) were incubated for at least 2.5 hours at 0°C before filtration over Whatman GF/B filters. Filters were additionally rinsed with 2 x 2 ml TrisHCl buffer (pH 7.4, 4°C). Filters were counted in Packard Emulsifier Safe scintillation fluid (3.5 ml) after 24 hours of extraction.
  • the tritiated antagonist [ 3 H]-1,3-dipropyl-8-cyclopentylxanthine ([ 3 H]DPCPX), and for the adenosine A 2A receptor, the tritiated antagonist [ 3 H]ZM 241385 were used. Since radiolabeled antagonists are not commercially available for the adenosine A3 receptor, [125I]AB-MECA, an A3 receptor agonist, was used. Displacement experiments were performed in the absence of GTP.
  • 2-iodoadenosine ( 1 ) was brominated at the 8-position prior to the introduction of the 2-substituent.
  • a standard bromination procedure e.g. stirring 2-iodoadenosine ( 1 ) or 2',3',5'-tri- O -acetyl-2-iodoadenosine ( 2 ) in dioxan and adding bromine water (in phosphate buffer, 10% w/v, pH ⁇ 7), 18 yielded the corresponding 8-bromo derivatives.
  • Table 2 displays radioligand binding data for all synthesized C2,8-adenosine derivatives (final products 1, 3-5, 7-23 ).
  • 8-Alkylamino adenosine derivatives had adenosine A 1 and A 2A receptor affinities in the ⁇ M range, without substantial preference for either receptor, whereas introduction of a cyclopentyl group at the N 6 -position led to an increase in adenosine A 1 receptor affinity up to 23-fold.
  • the 2-(1-hexynyl) adenosine derivatives 12-16 generally had higher adenosine A 2A receptor affinities than the 2-(( E )-1-hexenyl)-substituted compounds 17-21 , in line with the receptor affinities of compounds 3 and 4 .
  • the adenosine A 2A receptor affinity of compound 16 was higher than that of the 2-benzylamino-substituted derivative ( 23 ), also in line with receptor (functional) data on either 2-(1-hexynyl)- 8 or 2-benzylamino-substituted 20 derivatives.
  • Other adenosine derivatives substituted at the 8-position have not displayed very high receptor affinities.
  • 8-alkylamino-substituted adenosine derivatives were obtained with affinities in the low micromolar ( 17-20 ) or even nanomolar range (12-15) for the adenosine A 2A receptor.
  • the adenosine A 2A receptor affinity was somewhat decreased with the introduction of the 8-alkylamino groups, the A 2A receptor selectivity compared to (A 1 and) A 3 was increased.
  • Many 2-substituted adenosine derivatives have been described as potent ligands for the A 2A receptor, although binding data for the adenosine A 3 receptor is often lacking.
  • the selectivity for the adenosine A 2A receptor compared to A 3 was approx. 49- and 26-fold, respectively. Although the affinities of the 2-(1-hexenyl)adenosines were somewhat lower, the selectivity for the A 2A receptor was also increased.
  • the 8-substituents may have an influence on the conformation of the ligand itself (by forcing the ribose ring into the syn conformation).
  • the conformation of 8-bromoadenosine in the crystal structure is syn, suggested to cause the low affinity of this compound for the adenosine receptors.
  • the C2,8-disubstituted adenosine derivatives disclosed herein were synthesized in good overall yields starting from 2-iodoadenosine. Most compounds appear to have adenosine A 2A receptor affinities in the low micromolar or nanomolar range. Although the affinity for the adenosine A 2A receptor was decreased somewhat with the introduction of the 8-alkylamino substituents, the selectivity for this receptor compared to the A 3 receptor was improved significantly.
  • the adenosine A 2A receptor seem to accommodate the 8-propyl- or 8-butylamino substituents best, whereas the even larger 8-substituents (8-benzylamino or 8-(1-hexynyl)) were not well tolerated.
  • the 8-unsubstituted compounds 3 and 4 displayed a reduced intrinsic activity compared to the reference agonist CGS21680.
  • the intrinsic activity was either not affected much, or as in most cases, was further reduced, and most of the compounds behaved as partial agonists.
  • Most of the 8-substituted derivatives of 1 behaved as partial agonists as well.

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ATE532791T1 (de) * 2004-09-17 2011-11-15 Kissei Pharmaceutical In der 8-stellung modifiziertes purin- nukleosidderivat und dessen medizinische verwendung
GT200500281A (es) 2004-10-22 2006-04-24 Novartis Ag Compuestos organicos.
GB0500785D0 (en) 2005-01-14 2005-02-23 Novartis Ag Organic compounds
CN101410407A (zh) * 2006-04-03 2009-04-15 阿斯利康(瑞典)有限公司 取代腺嘌呤及其应用
ATE549337T1 (de) 2006-04-21 2012-03-15 Novartis Ag Purinderivate zur verwendung als adenosin-a2a- rezeptoragonisten
GB0607950D0 (en) 2006-04-21 2006-05-31 Novartis Ag Organic compounds
EP1889846A1 (en) 2006-07-13 2008-02-20 Novartis AG Purine derivatives as A2a agonists
EP1903044A1 (en) 2006-09-14 2008-03-26 Novartis AG Adenosine Derivatives as A2A Receptor Agonists
CN108129535A (zh) * 2017-12-19 2018-06-08 兰州奥凯化工公司 8-溴腺苷的合成方法
JP2024512077A (ja) * 2021-03-26 2024-03-18 フューチャー・メディシン・カンパニー・リミテッド Ａ２ａ及びａ３アデノシン受容体に拮抗作用をするアデノシン誘導体及びその製造方法

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WO2001040799A2 (en) * 1999-12-03 2001-06-07 Cv Therapeutics, Inc. Method of identifying partial adenosine a1 receptor agonists and their use in the treatment of arrhythmias
US6605597B1 (en) * 1999-12-03 2003-08-12 Cv Therapeutics, Inc. Partial or full A1agonists-N-6 heterocyclic 5′-thio substituted adenosine derivatives

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EP1368364A1 (en) 2003-12-10
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